Molecular studies of VZV infection, latency and reactivation in human neurons in-vitro

人类神经元水痘带状疱疹病毒感染、潜伏期和再激活的分子研究

基本信息

批准号：
9052861
负责人：
Paul R. Kinchington
金额：
$ 47.03万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-11-12 至 2020-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9052861
关键词：
Address Adult Afferent Neurons Age Animal Model Axon Blindness Capsid Chickenpox Climacteric Collaborations Communicable Diseases Defect Disease Effectiveness Event Eye Foundations Gene Deletion Gene Expression Genes Genetic Transcription Genomics Growth Herpes zoster disease Herpesviridae Herpesvirus Type 3 Homing Human Immune In Vitro Individual Infection Knowledge Libraries Life Light Lytic Modeling Molecular Nerve Neurologic Neurons Neurotropism Outcome Pain Peripheral Persons Prevention strategy Process Proteins Quality of life Recombinants Reporter Research Personnel Risk Role Site Small RNA Source Staging Stimulus System Temperature Testing Therapeutic Tissues Vaccines Viral Viral Genes Viral Genome Viral Proteins Virus Virus Diseases Virus Latency Virus Replication Work base chronic pain design genetic regulatory protein human embryonic stem cell improved mutant neuronal cell body next generation sequencing novel nuclease prevent programs prophylactic public health relevance reactivation from latency research study vector viral RNA virus genetics virus pathogenesis

项目摘要

DESCRIPTION (provided by applicant): There exists a great need to understand how the human herpesvirus Varicella zoster Virus (VZV) interacts with neurons. The sensory neuron is critical to successful VZV pathogenesis as the site of a decades-long state of persistence, and the source for VZV reactivation to cause Herpes zoster. This painful and debilitating disease is encountered by a third of adults, and is frequently complicated by chronic pain and neurological problems. Furthermore, ophthalmic zoster is a potentially devastating infectious disease that causes significant vision loss and a much greater toll than zoster elsewhere in terms of pain, complications and marked disruption of the quality of life. Even if all eligible persons received the zoster vaccine (which is far from being achieved). the partial effectiveness would still result in some half of a million zoster cases annually. What we know of VZV latency and reactivation has remained enigmatic, controversial and unclear, because most animal models or their neurological tissues do not support VZV replication or reactivation. Indeed, VZV experimental reactivation has proved exceedingly difficult under any circumstances. Yet, if neuronal infection, spread, persistence or reactivation can be prevented, disease could be more easily controlled. We now have an unprecedented cultured human neuron platform, derived from human embryonic stem cells, that supports not only VZV productive infection, but also persistent states that can now be reactivated. Our overlying hypothesis is that this system will enable us to address aspects of neuron infection, latency and qreactivation in a manner that has not been previously possible. Our first aim will test the hypothesis that specific VZV proteins are required for neuron axonal infection, replication, inter-neuronal spread or anterograde axonal return. We will evaluate fluorescent reporter VZV with gene deletions for each stage of neuronal infection to identify those proteins required. Such proteins could be not only be targeted in strategies to block latency and reactivation, but such VZV mutants could be the basis for improved vaccines with defined deficiencies in neurotropism. Our second aim will address the viral transcription program during VZV persistence and reactivation. We will then identify what transcription occurs during persistence and reactivation, including a search for small RNAs that may contribute to latency. We will also address if transcriptional programs differ following reactivation at differen temperatures. Our third aim will test the hypothesis that the persistent VZV genome can be targeted by large sequence recognition nucleases, in order to reduce VZV latent genomic loads or prevent reactivation. This would establish principles or targeting the latent VZV genome without reliance on viral gene products. Globally, our studies will set the stage for understanding and targeting the VZV latent state in a manner that has not been previously possible.

描述（通过应用程序证明）：存在一个非常需要了解urons的感觉，对于成功的VSV发病机理至关重要。在慢性疼痛和神经逻辑疾病中，眼科带状的疼痛是一种重要的视力损失，并且在带状疱疹的角色疫苗方面比带状疱疹更大在一半的带状障碍中，大多数动物模型或它们的神经组织都不支持VZV复制。现在，有一个未经培养的人神经元平台，该平台源自胚胎干细胞，不仅支持VZV生产感染，我们上覆的假设是Thastem，使我们能够以不具有特定的方式来解决感染，潜伏期和QREACTION的方面需要VZV蛋白对于神经元的轴突相关，我们将在神经元感染的每个阶段评估荧光报告的基因离子，以识别所需的蛋白质。在持久性和反应期间，将其定义为STHE病毒转录程序。 UR的第三个目标将假设可以通过大序列识别核酸酶来靶向帐篷基因组载荷或全球反应性。并以以前无法实现的方式针对VZV晚期。